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AAT Bioquest

FastClick™ XFD350 Alkyne

The reaction (Green Bar) of FastClick Cy5 Alkyne with coumarin azide occurs under extremely mild conditions (e.g., [Azide] = 0.02 mM, [Alkyne] = 0.02 mM, [CuSO4] = 0.02 mM, [Sodium Ascorbate] = 5 mM, in 100 mM HEPES) under which the common Cy5 alkyne does not effectively react with the coumarin azide substrate.
The reaction (Green Bar) of FastClick Cy5 Alkyne with coumarin azide occurs under extremely mild conditions (e.g., [Azide] = 0.02 mM, [Alkyne] = 0.02 mM, [CuSO4] = 0.02 mM, [Sodium Ascorbate] = 5 mM, in 100 mM HEPES) under which the common Cy5 alkyne does not effectively react with the coumarin azide substrate.
The reaction (Green Bar) of FastClick Cy5 Alkyne with coumarin azide occurs under extremely mild conditions (e.g., [Azide] = 0.02 mM, [Alkyne] = 0.02 mM, [CuSO4] = 0.02 mM, [Sodium Ascorbate] = 5 mM, in 100 mM HEPES) under which the common Cy5 alkyne does not effectively react with the coumarin azide substrate.
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Physical properties
Molecular weight621.71
SolventDMSO
Spectral properties
Correction Factor (260 nm)0.25
Correction Factor (280 nm)0.19
Extinction coefficient (cm -1 M -1)19000
Excitation (nm)343
Emission (nm)441
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure
Alternative formats
FastClick™ XFD350 Azide

OverviewpdfSDSpdfProtocol


See also: Click Chemistry
Molecular weight
621.71
Correction Factor (260 nm)
0.25
Correction Factor (280 nm)
0.19
Extinction coefficient (cm -1 M -1)
19000
Excitation (nm)
343
Emission (nm)
441
FastClick™ XFD350 Alkyne contains both the CAG moiety of FastClick (for assisting click efficiency) and Alexa Fluor® 350 fluorophore (as the fluorescence tag) for developing Alexa Fluor® 350-based fluorescent probes. FastClick™ XFD350 Alkyne readily reacts with an azido-modified biomolecule under extremely mild conditions. Alexa Fluor® 350 is a commonly used blue fluorophore for labeling proteins, nucleic acids, or other biomolecules. It has moderate photostability and excitation that matches the 350 nm laser line. Alexa Fluor® is a trademark of ThermoFisher Scientific. FastClick™ reagents have been developed by the scientists of AAT Bioquest for enhancing the yield and reaction speed of copper-catalyzed azide-alkyne cycloaddition (CuAAC) reaction. They contain a copper-chelating ligand that significantly stabilizes the Cu(I) oxidation state and thus accelerates the click reaction. They do not require the use of an external copper-chelator (such as the common THPTA or BTTAA). The high concentration of copper chelators is known to have a detrimental effect on DNA/RNA, thus causing biocompatibility issues. The introduction of a copper-chelating moiety at the reporter molecule allows for a dramatic raise of the effective Cu(I) concentration at the reaction site and thus accelerates the reaction. Under extremely mild conditions the FastClick™ azides and alkynes react much faster in high yield compared to the corresponding conventional CuAAC reactions. Click chemistry was developed by K. Barry Sharpless as a robust and specific method of ligating two molecules together. Two important characteristics make click chemistry attractive for assembling biomolecules. First, click reactions are bio-orthogonal, thus the click chemistry-functionalized biomolecules would not react with the natural biomolecules that lack a clickable functional group. Second, the reactions proceed with ease under mild conditions, such as at room temperature and in aqueous media.

Calculators


Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of FastClick™ XFD350 Alkyne to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

0.1 mg0.5 mg1 mg5 mg10 mg
1 mM160.847 µL804.233 µL1.608 mL8.042 mL16.085 mL
5 mM32.169 µL160.847 µL321.693 µL1.608 mL3.217 mL
10 mM16.085 µL80.423 µL160.847 µL804.233 µL1.608 mL

Molarity calculator

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Mass (Calculate)Molecular weightVolume (Calculate)Concentration (Calculate)Moles
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Spectrum


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spectrum

Spectral properties

Correction Factor (260 nm)0.25
Correction Factor (280 nm)0.19
Extinction coefficient (cm -1 M -1)19000
Excitation (nm)343
Emission (nm)441

Product Family


NameExcitation (nm)Emission (nm)Extinction coefficient (cm -1 M -1)Quantum yieldCorrection Factor (260 nm)Correction Factor (280 nm)
FastClick™ Cy3 Alkyne55556915000010.1510.070.073
FastClick™ Cy5 Alkyne65167025000010.271, 0.420.020.03
FastClick™ Cy7 Alkyne7567792500000.30.050.036
FastClick™ XFD488 Alkyne499520710000.9210.300.11
FastClick™ XFD555 Alkyne5535681500000.110.080.08
FastClick™ XFD647 Alkyne6506712390000.3310.000.03
FastClick™ XFD750 Alkyne7527762400000.1210.000.04

Images


References


View all 50 references: Citation Explorer
Click chemistry approaches for developing carbonic anhydrase inhibitors and their applications.
Authors: Angeli, Andrea and Supuran, Claudiu T
Journal: Journal of enzyme inhibition and medicinal chemistry (2023): 2166503
Polydopamine nanoparticle-mediated, click chemistry triggered, microparticle-counting immunosensor for the sensitive detection of ochratoxin A.
Authors: Chen, Rui and Dong, Yongzhen and Hong, Feng and Zhang, Xiya and Wang, Xiaohong and Wang, Jia and Chen, Yiping
Journal: Journal of hazardous materials (2022): 128206
Design, Synthesis and Biological Evaluation of New Carbohydrate-Based Coumarin Derivatives as Selective Carbonic Anhydrase IX Inhibitors via "Click" Reaction.
Authors: Chu, Naying and Wang, Yitong and Jia, Hao and Han, Jie and Wang, Xiaoyi and Hou, Zhuang
Journal: Molecules (Basel, Switzerland) (2022)
Late-Stage Functionalization through Click Chemistry Provides GLUT5-Targeting Glycoconjugate as a Potential PET Imaging Probe.
Authors: Oronova, Adelina and Tanasova, Marina
Journal: International journal of molecular sciences (2022)
New Azido Coumarins as Potential Agents for Fluorescent Labeling and Their "Click" Chemistry Reactions for the Conjugation with closo-Dodecaborate Anion.
Authors: Laskova, Julia and Serdyukov, Alexander and Kosenko, Irina and Ananyev, Ivan and Titova, Ekaterina and Druzina, Anna and Sivaev, Igor and Antonets, Anastasia A and Nazarov, Alexey A and Bregadze, Vladimir I
Journal: Molecules (Basel, Switzerland) (2022)
Chemoselective bioconjugation based on modular click chemistry with 4-halocoumarins and aryl sulfonates.
Authors: Thanzeel, F Yushra and Wolf, Christian
Journal: RSC advances (2021): 18960-18965
Click Chemistry-Assisted Bioconjugates for Hapten Immunodiagnostics.
Authors: López-Puertollano, Daniel and Agulló, Consuelo and Mercader, Josep V and Abad-Somovilla, Antonio and Abad-Fuentes, Antonio
Journal: Bioconjugate chemistry (2020): 956-964
Click chemistry based regioselective one-pot synthesis of coumarin-3-yl-methyl-1,2,3-triazolyl-1,2,4-triazol-3(4H)-ones as newer potent antitubercular agents.
Authors: Somagond, Shilpa M and Kamble, Ravindra R and Bayannavar, Praveen K and Shaikh, Saba Kauser J and Joshi, Shrinivas D and Kumbar, Vijay M and Nesaragi, Aravind R and Kariduraganavar, Mahadevappa Y
Journal: Archiv der Pharmazie (2019): e1900013
DNA template-mediated click chemistry-based portable signal-on sensor for ochratoxin A detection.
Authors: Qiu, Suyan and Yuan, Lijuan and Wei, Yihua and Zhang, Dawen and Chen, Qinglong and Lin, Zhenyu and Luo, Linguang
Journal: Food chemistry (2019): 124929
Tag-Convertible Photocrosslinker with Click-On/Off N-Acylsulfonamide Linkage for Protein Identification.
Authors: Hayashi, Ryuji and Morimoto, Shota and Tomohiro, Takenori
Journal: Chemistry, an Asian journal (2019): 3145-3148